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Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay
Chen, Huaxin1,2,3; Jiang, Peng1,2,3
2018-12-01
Source PublicationJOURNAL OF BIOSCIENCE AND BIOENGINEERING
ISSN1389-1723
Volume126Issue:6Pages:778-782
Corresponding AuthorChen, Huaxin(chenhx001@126.com)
AbstractFusion protein of streptavidin and allophycocyanin holo-alpha subunit (ApcA) is fluorescent and is able to bind biotin. This fusion protein (SLA) can be used as fluorescent label in immunofluorescence assay. In this study, one or two repeats of ApcA were fused to the protein SLA, with the aim to improve its brightness. The fusion proteins SLA2 (two repeats of ApcA) and SLA3 (three repeats of ApcA), together with lyase (cpcS) and phycoerythrobilin synthesizing enzymes (Ho1 and PebS), were co-expressed in Escherichia coli. These fusion proteins were purified by affinity chromatography. While SLA2 and SLA3 shared similar absorbance spectra, fluorescence spectra and biotin-binding activities with SLA, their brightness were much higher than that of SLA. When used as fluorescent labels in immunofluorescence assay, SLA2 and SLA3 showed higher detection sensitivity than SLA. These results suggested that SLA2 and SLA3 were the preferable fluorescent labels in immunofluorescence assays. (C) 2018, The Society for Biotechnology, Japan. All rights reserved.
KeywordAllophycocyanin Streptavidin Fusion protein Fluorescence Immunofluorescence assay
DOI10.1016/j.jbiosc.2018.06.004
Indexed BySCI
Language英语
Funding ProjectDevelopment Funds of Science and Technology of Shinan District[2016-3-010-ZH] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDA11030202] ; National 863 Program[2014AA093505] ; National 863 Program[2014AA093501]
WOS Research AreaBiotechnology & Applied Microbiology ; Food Science & Technology
WOS SubjectBiotechnology & Applied Microbiology ; Food Science & Technology
WOS IDWOS:000452933700017
PublisherSOC BIOSCIENCE BIOENGINEERING JAPAN
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Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/155791
Collection实验海洋生物学重点实验室
Corresponding AuthorChen, Huaxin
Affiliation1.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266071, Peoples R China
3.Chinese Acad Sci, Ctr Ocean Mega Sci, Qingdao 266071, Peoples R China
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Corresponding Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Chen, Huaxin,Jiang, Peng. Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay[J]. JOURNAL OF BIOSCIENCE AND BIOENGINEERING,2018,126(6):778-782.
APA Chen, Huaxin,&Jiang, Peng.(2018).Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay.JOURNAL OF BIOSCIENCE AND BIOENGINEERING,126(6),778-782.
MLA Chen, Huaxin,et al."Combinational biosynthesis and characterization of fusion proteins with tandem repeats of allophycocyanin holo-alpha subunits, and their application as bright fluorescent labels for immunofluorescence assay".JOURNAL OF BIOSCIENCE AND BIOENGINEERING 126.6(2018):778-782.
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